Benzovindiflupyr at estimated drinking water concentrations induces hepatic injury via the NF-κB signaling pathway: In vivo, in vitro, and in silico studies.

While the extensive application of benzovindiflupyr (BZF) contributes to crop protection, it may pose potential threats to non-target species. However, limited research has investigated the effects of environmentally relevant concentrations of BZF on non-target organisms. By integrating in vivo, in vitro, and in silico approaches, we revealed that BZF induces hepatic injury in zebrafish via the NF-κB signaling pathway activation at estimated drinking water concentrations. Specifically, following a 28-day exposure to gradient BZF concentrations (0, 5 and 50 μg/L), adult zebrafish exhibited marked hepatotoxicity. Based on transcriptomic data, we deduced that the NF-κB signaling pathway was involved. Subsequent experiments in HepG2 cells confirmed that the NF-κB inhibitor PDTC attenuated NF-κB-related transcriptional and protein responses, verifying the pathway's critical role. Furthermore, molecular docking analysis provided supportive structural evidence for the binding of BZF to NF-κB-related proteins. Overall, through a multi-model approach, we confirmed that the NF-κB signaling pathway is a crucial mediator of hepatic injury in zebrafish following BZF exposure. These findings elucidate the mechanism of BZF toxicity and underscore the potential ecological threats to aquatic organisms under realistic exposure scenarios.